Insights for producing and future use of induced pluripotent stem cells from the small red brocket deer (Mazama jucunda)

Abstract

The small red brocket (Mazama jucunda), a deer endemic to the Atlantic Forest biome, is considered vulnerable by the IUCN, needing reproductive biotechnologies to preserve its genetic material. In this context, this study aimed to develop a proof-of-concept protocol for producing and characterizing induced pluripotent stem cells (iPSCs) originated from adult Mazama jucunda fibroblasts. Fibroblasts from skin biopsies were isolated and characterized regarding morphology and doubling time. Two transduction experiments were carried out with three technical replicates: transduction of eGFP reporter gene and transduction of pluripotency factors (murine OSKM - OCT4, SOX2, KLF4 and c-MYC) aiming for cellular reprogramming. This study successfully produced GFP+ cell lines, validating the transduction capacity of the lentiviral delivery system, as well as establishing a GFP+ cell line from the small red brocket, used for the OSKM transduction. In the OSKM transduction, cell aggregates with typical colony morphology were observed 33 days after the first transduction, however the colonies did not maintain classical characteristics in culture. Obtaining iPSCs from somatic cells is a very promising biotechnology and, currently, there has only been one report of iPSC induction in a cervid species, making studies such as these great contributions for better understanding reprogramming pathways in different deer species for future use in conservation programs.